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2021 ◽  
Vol 11 (3) ◽  
pp. 366-372
Author(s):  
Chaochao Zhong ◽  
Xinchong Huang ◽  
Yibin Qin ◽  
Haihang Miao ◽  
Weiwei Zhou

To explore the influence and mechanism of lncRNA SNHG14 on oxidized low-density lipoprotein (oxLDL)-induced vascular endothelial cell injury, cellular levels of SNHG14 and miRNA-93-5p were detected in oxLDL-induced vascular endothelial cells by RT-qPCR, and the levels of MDA, SOD and GSH-Px were detected by ELISA. Flow cytometry was used to detect cellular apoptosis, and western blot analysis was used to determine the abundance of Bcl-2 and Bax proteins. SNHG14 small interfering RNA or miRNA-93-5p mimics were transfected into vascular endothelial cells to interfere with SNHG14 expression or overexpress miRNA-93-5p. To study the effects of interfering with SNHG14 expression or overexpression of miRNA-93-5p, the levels of MDA, SOD and GSH-Px, apoptosis and the levels of Bcl-2 and Bax protein were studied in oxLDL-induced endothelial cells with either altered SNHG14 expression or overexpressed miRNA-93-5p. A dual-luciferase reporter gene experiment verified the regulatory connection between SNHG14 and miRNA-93-5p. After oxLDL acted on vascular endothelial cells, the expression levels of SNHG14 were significantly increased, while the expression levels of miRNA-93-5p were significantly reduced, MDA levels were increased, and SOD and GSH-Px activities were reduced. Both the apoptosis rate and Bax protein levels were significantly increased, and Bcl-2 expression was reduced. Interference with SNHG14 expression or overexpression of miRNA-93-5p can reduce the MDA content in oxLDL-induced vascular endothelial cells, increase the activity of SOD and GSH-Px, and reduce the apoptosis rate and Bax protein levels, and promote Bcl-2 expression. SNHG14 targeted to negatively regulate miRNA-93-5p expression, inhibited miRNA-93-5p expression and reversed the interference of SNHG14 expression with oxLDL-induced variation in MDA, SOD and GSH-Px levels, apoptosis rate, and Bax and Bcl-2 protein levels. Interference of SNHG14 expression may reduce oxidative stress and apoptosis of vascular endothelial cells induced by oxLDL by negatively regulating the expression of miRNA-93-5p.


Diagnostics ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 868
Author(s):  
Workneh Korma ◽  
Adane Mihret ◽  
Azeb Tarekegn ◽  
Yunhee Chang ◽  
Dasom Hwang ◽  
...  

The diagnosis and prognosis of tuberculosis remains challenging and necessitates the development of a new test that can accurately diagnose and monitor treatment responses. In this regard, miRNA is becoming a potential diagnostic and prognostic biomarker which differentiates treatment respondents from non-respondents for various non-infectious and infectious diseases, including tuberculosis. The concentration of miRNAs varies based on cell type, disease, and site of infection, implicating that selection of an optimal reference gene is crucial, and determines the quantification of transcript level and biological interpretation of the data. Thus, the study evaluated the stability and expression level of five candidate miRNAs (let-7i-5p, let-7a-5p, miRNA-16-5p, miRNA-22-3p and miRNA-93-5p), including U6 Small Nuclear RNA (RNU6B) to normalize circulating miRNAs in the plasma of 68 participants (26 healthy controls, 23 latent, and 19 pulmonary tuberculosis infected) recruited from four health centers and three hospitals in Addis Ababa, Ethiopia. The expression levels of miRNAs isolated from plasma of culture confirmed newly diagnosed pulmonary tuberculosis patients were compared with latently infected and non-infected healthy controls. The qPCR data were analyzed using four independent statistical tools: Best Keeper, Genorm, Normfinder and comparative delta-Ct methods, and the data showed that miRNA-22-3p and miRNA-93-5p were suitable plasma reference miRNAs in a tuberculosis study.


2020 ◽  
Vol 40 (2) ◽  
Author(s):  
Liu Lan ◽  
Zhishan Liang ◽  
Yingxi Zhao ◽  
Yuzhen Mo

Abstract Background: MCM3AP antisense RNA 1 (MCM3AP-AS1) is characterized as an oncogenic lncRNA in hepatocellular carcinoma and glioblastoma. We analyzed TCGA dataset and observed the down-regulation of MCM3AP-AS1 in cervical squamous cell carcinoma (CSCC). The present study was therefore performed to investigate the role of MCM3AP-AS1 in CSCC. Methods: A total of 64 female patients with CSCC (38–68 years old; mean age: 53.1 ± 6.5 years old) were enrolled in the present study. RT-qPCR was performed to evaluate gene expression. Methylation specific PCR (MSP) was performed to assess the methylation of miR-93 gene after the overexpression and silencing of MCM3AP-AS1. Cell transfections were performed to investigate the interactions between MCM3AP-AS1 and miR-93. Cell proliferation was assessed by CCK-8 assay. Results: The results showed that MCM3AP-AS1 was down-regulated in CSCC and predicted poor survival. The expression levels of MCM3AP-AS1 were inversely correlated with the expression levels of miR-93. Overexpression of MCM3AP-AS1 led to down-regulation of miR-93, while silencing of MCM3AP-AS1 played an opposite role in CSCC cells. Methylation-specific PCR revealed that MCM3AP-AS1 could positively regulate the methylation of miR-93 gene. Cell proliferation analysis showed that overexpression of MCM3AP-AS1 led to reduced proliferation rate of CSCC cells. Silencing of MCM3AP-AS1 played an opposite role and overexpression of miR-93 reduced the effects of overexpressing MCM3AP-AS1. Conclusions: Therefore, MCM3AP-AS1 may inhibit cell proliferation in CSCC by down-regulating miRNA-93.


2020 ◽  
Vol 32 (1) ◽  
pp. 1111-1115 ◽  
Author(s):  
Wei Zheng ◽  
Jianying Yang ◽  
Yuhong Wang ◽  
Xinru Liu

2019 ◽  
Vol 20 (22) ◽  
pp. 5750 ◽  
Author(s):  
Zofia Ulańczyk ◽  
Anna Sobuś ◽  
Karolina Łuczkowska ◽  
Aleksandra Grabowicz ◽  
Katarzyna Mozolewska-Piotrowska ◽  
...  

Age-related macular degeneration (AMD) remains the leading cause of blindness in elderly people, but the pathophysiology of this disease is still largely unknown. We investigated the systemic expression of angiogenesis-regulating growth factors and selected miRNAs known to regulate angiogenesis in AMD patients. We also focused on possible correlations of their expression with the presence of CFH Y402H or ARMS A69S risk variants. A total of 354 AMD patients and 121 controls were enrolled in this study. The levels of angiogenesis-regulating factors were analyzed in plasma samples using Luminex technology. The expression of selected miRNAs was analyzed in peripheral blood plasma using real-time qPCR. The genetic analysis was performed with an Illumina NextSeq500 system. AMD was an independent factor associated with lower levels of angiogenin (β = −0.29, p < 0.001), endostatin (β = −0.18, p < 0.001), FGF-basic (β = −0.18, p < 0.001), PlGF (β = −0.24, p < 0.001), miRNA-21-3p (β = −0.13, p = 0.01) and miRNA-155-5p (β = −0.16, p = 0.002); and with higher levels of FGF-acidic (β = 0.11, p = 0.03), miRNA-23a-3p (β = 0.17, p < 0.001), miRNA-126-5p (β = 0.13, p = 0.009), miRNA-16-5p (β = 0.40, p < 0.001), miRNA-17-3p (β = 0.13, p = 0.01), miRNA-17-5p (β = 0.17, p < 0.001), miRNA-223-3p (β = 0.15, p = 0.004), and miRNA-93 (β = 0.11, p = 0.04). The expression of analyzed miRNA molecules significantly correlated with the levels of tested angiogenesis-regulating factors and clinical parameters in AMD patients, whereas such correlations were not observed in controls. We also found an association between the CFH Y402H polymorphism and miRNA profiles, whereby TT homozygotes showed evidently higher expression of miRNA-16-5p than CC homozygotes or TC heterozygotes (p = 0.0007). Our results suggest that the balance between systemic pro- and anti-angiogenic factors and miRNAs is vital in multifactorial AMD pathogenesis.


2019 ◽  
Vol 240 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Zerui Wu ◽  
Lin Cai ◽  
Jianglong Lu ◽  
Chengde Wang ◽  
Jiaqing Guan ◽  
...  

To date, the management of dopamine agonist (DA)-resistant prolactinomas remains a major clinical problem. Previously, we determined that miRNA-93 expression increases in DA-resistant prolactinomas; however, the role of miRNA-93 in the DA resistance remains largely unexplored. Hence, this study aimed to investigate the susceptibility of tumor cells to cabergoline (CAB) and the autophagy changes in MMQ and GH3 cells after miRNA-93 overexpression or inhibition. We used bioinformatics to identify the potential target of miRNA-93. Subsequently, we analyzed the correlation between miRNA-93 and autophagy-related 7 (ATG7) using protein expression analysis and luciferase assays. Furthermore, the change in the effect of miRNA-93 was measured after ATG7 overexpression. miRNA-93 expression was elevated in DA-resistant prolactinomas, whereas the expression of its identified target, ATG7, was downregulated. miRNA-93 overexpression suppressed the cytotoxic effect of CAB in MMQ and GH3 cells. In contrast, miRNA-93 downregulation enhanced CAB efficiency and promoted cell autophagy, eventually resulting in apoptosis. These results were further confirmed in in vivo xenograft models in nude mice. ATG7 overexpression could reverse the inhibitory effect of miRNA-93 on CAB treatment. Taken together, our results suggest that miRNA-93 mediates CAB resistance via autophagy downregulation by targeting ATG7 and serves as a promising therapeutic target for prolactinoma.


Tumor Biology ◽  
2018 ◽  
Vol 40 (5) ◽  
pp. 101042831877586 ◽  
Author(s):  
Ahmed Hussein Zedan ◽  
Torben Frøstrup Hansen ◽  
Jannie Assenholt ◽  
Mindaugas Pleckaitis ◽  
Jonna Skov Madsen ◽  
...  

Prostate cancer is the most common cancer among men in the western world. Clinical practice is continuously challenged by the pitfalls of the available diagnostic tools. microRNAs may represent promising biomarkers in many types of human cancers, including prostate cancer. The aim of this study was to investigate microRNA expression in tumour tissue and matched plasma in a cohort of patients with primary metastatic prostate cancer. The relative expression of 12 microRNAs was assessed in diagnostic needle biopsies from the prostate and matched plasma samples in two prospective cohorts (screening cohorts) comprising 21 patients with metastatic prostate cancer and 25 control patients. An independent validation cohort of plasma samples was collected prospectively from 149 newly diagnosed patients with local/locally advanced prostate cancer. Analyses were performed using real-time polymerase chain reaction. miRNA-93 showed a significant negative correlation between expression in tumour tissue and plasma in patients with metastatic prostate cancer. Furthermore, the plasma level of miRNA-93 significantly decreased after treatment in patients with local/locally advanced prostate cancer compared to baseline plasma level. The expression of six microRNAs (let-7b, miRNA-34a, -125b, -143, -145 and -221) was downregulated, and three microRNAs (miRNA-21, -25 and miRNA-93) were upregulated in tumour tissue compared to benign prostate tissue. In plasma, six microRNAs were upregulated (miRNA-21, -125b, -126, -141, -143 and -375), while let-7b was downregulated in patients with metastatic prostate cancer compared to the control cohort. In the metastatic prostate cancer cohort, the expression of four microRNAs (miRNA-125b, -126, -143 and -221), and miRNA-141 in tissue was associated with Gleason score and prostate-specific antigen, respectively. The expression of miRNA-93 in tumour tissue was correlated with matched plasma levels and showed a significant decrease in plasma level after intervention in local prostate cancer. Differential expression between tumour and benign prostate was detected for several microRNAs in both tissue and plasma.


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